Methods using dsdna to mediate rna interference (rnai)

Details Methods using dsdna to mediate rna interference (rnai) Methods using dsdna to mediate rna interference (rnai)

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) C12N 15/00 (2006.01) C12N 15/11 (2006.01)

Patent

CA 2501065

The present invention provides methods of producing dsDNA molecules that can be used to mediate RNA interference (RNAi). These methods include the production of hairpin DNAs including random sequences, and the use of convergent promoters to co-express sense and antisense RNAs. As such, the invention allows the production of random short hairpin RNA (shRNA) and amsll interfering RNA (siRNA) expression libraries for forward genetic screening.

L'invention concerne des procédés de production de molécules d'ADNds qui peuvent servir dans la médiation des ARN interférents (ARNi). Ces procédés consistent à préparer des ADN ayant des structures en épingle à cheveux qui comportent des séquences aléatoires, et à utiliser des promoteurs convergents pour coexprimer des ARN sens et anti-sens. L'invention permet ainsi d'obtenir des banques d'expression d'ARN courts en épingle à cheveux aléatoires (shRNA) et de petits ARN interférents (siRNA) qui seront utilisées dans le criblage génétique.

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